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Summary: We have identified a novel gene designated CsV03-3 from Citrus sinensis (L.) Osbeck that encodes a 50 amino acid polypeptide with a predicted molecular mass of 5.4kDa and a pI of 3.7. CsV03-3 expression is up-regulated by application of methyl jasmonate, salicylic acid, and abscisic acid as well as by abiotic stress and insect herbivory. CsV03-3 belongs to a small gene family consisting of at least three other closely related members (CsV03-1, CsV03-2, and CsV03-4) whose expression is also responsive to phytohormone application and abiotic stress. Sequence similarity searches of the public databases were unsuccessful in finding sequence homologs to CsV03-3 or any CsV03 family member; however, structural prediction models coupled with model comparison to Protein Data Bank folds indicated that the predicted polypeptide encoded by CsV03-3 has structural similarity to proteins with nucleic acid binding activity. Gel mobility shift assays performed on recombinant CsV03-3 protein demonstrated active binding with dsDNA and, to a lesser extent, ssDNA. Based on the phytohormone-inducible expression patterns, the ability to bind nucleic acids, and the lack of significant sequence homology to public databases, we propose that CsV03-3 and its related homologs defines a new class of nucleic acid binding proteins that are responsive to defense and stress signaling in woody perennials. [Copyright &y& Elsevier]

Abstract: The molecular mechanisms underlying plant defense to sap-feeding insects are slowly being uncovered. In large part, past research has focused on interactions between phloem-feeding insects and their annual host plants with little emphasis on xylem-feeders or woody perennials—especially fruit trees. Using nylon filter cDNA arrays, we analyzed the transcriptional changes of 1731 non-redundant citrus transcripts that resulted from herbivory by a xylem-feeding leafhopper, the glassy-winged sharpshooter (GWSS), Homalodisca coagulata (Say) (Hemiptera: Cicadellidae). In addition, herbivory-elicited changes were compared to those of mechanical damage to better identify GWSS-specific responses. GWSS feeding led to a significant expression change in 50 transcripts. Of these, 14 were also changed by mechanical damage; however, the magnitude was in many cases reduced, suggesting transcriptional modification by GWSS-derived elicitors. Sequence similarity searches with the public database GenBank indicated that the responsive transcripts broadly function in direct defense, defense signaling, ROS scavenging, transport, cell wall modification, photosynthesis and abiotic stress. In particular, GWSS feeding resulted in a transcript profile that resembled wounding, likely through jasmonic acid-independent pathways, as well as an association with dehydration stress. In contrast to similar studies with aphids, salicylic acid-dependent pathogenesis-elated genes were weakly induced. Interestingly, six of the GWSS-responsive transcripts failed to significantly match any public protein sequence signifying their potential as novel genes functioning in plant defense, wound response or abiotic stress. [Copyright &y& Elsevier]